Mesenchymal stem cells (MSCs) contained in bone marrow fluid, and the like, have multi-differentiation potency to differentiate into a variety of cells such as an osteocyte, a chondrocyte, an adipocyte, a myocyte, a stromal cell, a neurocyte, and a tendon cell, thus attracting attention as a cell source for cell therapy and regenerative medicine. However, only a small amount of mesenchymal stem cells can be collected from the bone marrow fluid and the like. Therefore, for use in clinical treatment, it is important that mesenchymal stem cells collected from bone marrow fluid are isolated following concentration, and proliferated to a large amount in a short time.
As a method for efficiently proliferating mesenchymal stem cells, for example, methods disclosed in Patent Document 1 and Patent Document 2 are known.
In these methods, a fibroblast growth factor is added to a medium as a proliferation stimulant for mesenchymal stem cells.
Moreover, fetal bovine serum or human serum is typically used for a medium used for culturing mesenchymal stem cells.
Moreover, as a method for culturing mesenchymal stem cells in bone marrow fluid after the concentration/isolation thereof, a method is known in which, following centrifugation of the collected bone marrow fluid, the supernatant liquid is removed, the remaining precipitate portion alone is seeded in a culture vessel, and the mesenchymal stem cells are proliferated while they are adhered to the bottom face. In this case, a method for cell culture is proposed in which the seeding density of the mesenchymal stem cells adhered to the bottom face of the culture vessel is adjusted to an appropriate value (for example, refer to Patent Document 3).